期刊
JOURNAL OF CELL SCIENCE
卷 134, 期 19, 页码 -出版社
COMPANY BIOLOGISTS LTD
DOI: 10.1242/jcs.258409
关键词
Epidermolysis bullosa simplex; EBS; Keratin mutation; EGFR signalling; ERK1/2 signalling; EGFR inhibitors; Pathway intervention therapy drug screen
类别
资金
- DEBRA International grants [LANE2/LANE3, IAF311011, SPF2013/004]
- Biomedical Research Council of Singapore
In Epidermolysis Bullosa Simplex (EBS), mutations in keratin 14 or keratin 5 can lead to skin blistering due to keratinocyte rupture. A semi-automated system was developed to quantify the effects of test compounds on keratin aggregates, revealing potential candidates that can reduce aggregates and impact signaling pathways. Additionally, exposure to EGF ligand induced keratin aggregates, but this effect was reversible with EGFR inhibition, suggesting potential therapeutic avenues for reducing skin fragility in EBS patients.
In the skin fragility disorder epidermolysis bullosa simplex (EBS), mutations in keratin 14 (K14, also known as KRT14) or keratin 5 (K5, also known as KRT5) lead to keratinocyte rupture and skin blistering. Severe forms of EBS are associated with cytoplasmic protein aggregates, with elevated kinase activation of ERK1 and ERK2 (ERK1/2; also known as MAPK3 and MAPK1, respectively), suggesting intrinsic stress caused by misfolded keratin protein. Human keratinocyte EBS reporter cells stably expressing GFP-tagged EBS-mimetic mutant K14 were used to optimize a semiautomated system to quantify the effects of test compounds on keratin aggregates. Screening of a protein kinase inhibitor library identified several candidates that reduced aggregates and impacted on epidermal growth factor receptor (EGFR) signalling. EGF ligand exposure induced keratin aggregates in EBS reporter keratinocytes, which was reversible by EGFR inhibition. EBS keratinocytes treated with a known EGFR inhibitor, afatinib, were driven out of activation and towards quiescence with minimal cell death. Aggregate reduction was accompanied by denser keratin filament networks with enhanced intercellular cohesion and resilience, which when extrapolated to a whole tissue context would predict reduced epidermal fragility in EBS patients. This assay system provides a powerful tool for discovery and development of new pathway intervention therapeutic avenues for EBS.
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